Abstract：
BACKGROUND: Mounting experimental evidence has underscored the remarkable role played by the Wnt family of proteins in the spinal cord functioning and therapeutic potential in spinal cord injury (SCI). We aim to provide a therapeutic prospect associated with the modulation of canonical Wnt signaling, examining the spatio-temporal expression pattern of Dickkopf-1 (Dkk1) and its neutralization after SCI. We employ an intraparenchymal injection of the clinically validated Dkk1-blocking antibody, BHQ880, to elucidate its effects in SCI.
METHODS: A rat model of contusion SCI was used. Histological analyses were performed, wherein Dkk1 protein was sought, and ELISA analyses were employed for Dkk1 detection in cerebrospinal fluid and serum. To ascertain the BHQ880 therapeutic effect, rats were subjected to SCI and then injected with the antibody in the lesion epicenter 24 hours post-injury (hpi). Subsequent evaluation of motor functional recovery extended up to 56 days post-injury (dpi). qRT-PCR and histological analyses were conducted.
RESULTS: We demonstrate the presence of Dkk1 in the healthy rat spinal cord, with pronounced alterations observed following injury, primarily concentrated in the epicenter regions. Notably, a significative upregulation of Dkk1 was detected at 24 hpi, peaking at 3 dpi and remaining elevated until 42 dpi. Moreover, we revealed that early administration of BHQ880 considerably improved motor functional recovery, promoted preservation of myelinated tissue, and reduced astroglial and microglia/macrophage reactivity. Furthermore, there was a decrease in the acute expression of different inflammatory genes.
CONCLUSIONS: Collectively, our findings highlight the therapeutic potential of BHQ880 treatment in the context of SCI.
METHODS: A rat model of contusion SCI was used. Histological analyses were performed, wherein Dkk1 protein was sought, and ELISA analyses were employed for Dkk1 detection in cerebrospinal fluid and serum. To ascertain the BHQ880 therapeutic effect, rats were subjected to SCI and then injected with the antibody in the lesion epicenter 24 hours post-injury (hpi). Subsequent evaluation of motor functional recovery extended up to 56 days post-injury (dpi). qRT-PCR and histological analyses were conducted.
RESULTS: We demonstrate the presence of Dkk1 in the healthy rat spinal cord, with pronounced alterations observed following injury, primarily concentrated in the epicenter regions. Notably, a significative upregulation of Dkk1 was detected at 24 hpi, peaking at 3 dpi and remaining elevated until 42 dpi. Moreover, we revealed that early administration of BHQ880 considerably improved motor functional recovery, promoted preservation of myelinated tissue, and reduced astroglial and microglia/macrophage reactivity. Furthermore, there was a decrease in the acute expression of different inflammatory genes.
CONCLUSIONS: Collectively, our findings highlight the therapeutic potential of BHQ880 treatment in the context of SCI.
摘要：
背景：大量的实验证据强调了Wnt蛋白家族在脊髓损伤（SCI）的脊髓功能和治疗潜力中的显着作用。我们的目标是提供与经典Wnt信号调节相关的治疗前景，检查Dickkopf-1（Dkk1）的时空表达模式及其在SCI后的中和。我们使用临床验证的Dkk1阻断抗体的实质内注射，BHQ880，以阐明其在SCI中的作用。
方法：采用大鼠脊髓损伤模型。进行了组织学分析，其中寻找Dkk1蛋白，和ELISA分析用于脑脊液和血清中的Dkk1检测。为了确定BHQ880的治疗效果，对大鼠进行SCI,然后在损伤后24小时(hpi)在损伤中心注射抗体。随后对运动功能恢复的评估持续到受伤后（dpi）的56天。进行qRT-PCR和组织学分析。
结果：我们证明了健康大鼠脊髓中Dkk1的存在，损伤后观察到明显的变化，主要集中在震中地区。值得注意的是,在24hpi时检测到Dkk1的显着上调，在3dpi达到峰值，并保持升高直到42dpi。此外，我们发现BHQ880的早期给药显著改善了运动功能恢复，促进髓鞘组织的保存，并降低星形胶质细胞和小胶质细胞/巨噬细胞的反应性。此外，不同炎症基因的急性表达减少。
结论：总的来说，我们的研究结果强调了BHQ880治疗在SCI背景下的治疗潜力.
方法：采用大鼠脊髓损伤模型。进行了组织学分析，其中寻找Dkk1蛋白，和ELISA分析用于脑脊液和血清中的Dkk1检测。为了确定BHQ880的治疗效果，对大鼠进行SCI,然后在损伤后24小时(hpi)在损伤中心注射抗体。随后对运动功能恢复的评估持续到受伤后（dpi）的56天。进行qRT-PCR和组织学分析。
结果：我们证明了健康大鼠脊髓中Dkk1的存在，损伤后观察到明显的变化，主要集中在震中地区。值得注意的是,在24hpi时检测到Dkk1的显着上调，在3dpi达到峰值，并保持升高直到42dpi。此外，我们发现BHQ880的早期给药显著改善了运动功能恢复，促进髓鞘组织的保存，并降低星形胶质细胞和小胶质细胞/巨噬细胞的反应性。此外，不同炎症基因的急性表达减少。
结论：总的来说，我们的研究结果强调了BHQ880治疗在SCI背景下的治疗潜力.
